Samples of rectal faeces were collected immediately after slaughter from 400 cattle each month for a 1-year period and from 1000 each of sheep, pigs and poultry over the same period. Samples were examined for Escherichia coli O157 by enrichment culture in buffered peptone water with vancomycin, cefixime and cefsulodin followed by immunomagnetic separation and culture of magnetic particles onto cefixime tellurite sorbitol MacConkey agar. E. coli O157 was isolated from 752 (15.7%) of 4800 cattle, 22 (2.2%) of 1000 sheep and from 4 (0.4%) of 1000 pigs, but not from any of 1000 chickens. Of the cattle sampled. 1840 (38.4%) were prime beef animals, 1661 (34.6%) were dairy animals being culled and the status could not be determined for the other 1299 (27%) animals. E. coli O157 was found in 246 (13.4%) of the 1840 beef cattle and 268 (16.1%) of the 1661 dairy cattle. The monthly prevalence of E. coli O157 in cattle was 4.8-36.8% and was at its highest in spring and late summer. Seventeen of the 22 isolates from sheep were also made over the summer period. All E. coli O157 isolates from sheep and 749 (99.6%) of the 752 E. coli O157 isolates from cattle were verocytotoxigenic as determined by Vero cell assay and DNA hybridization, eaeA gene positive, contained a 92 kb plasmid and were thus typical of strains causing infections in man. In contrast isolates from pigs were non-toxigenic, eaeA gene negative and did not contain a 92 kb plasmid and would, therefore, be unlikely to be a source of infection for man.

SUMMARYIn May-June 1992 cases of infection with verocytotoxin-producing (VT+) Escherichia coli 0157 in South Yorkshire could have been associated with prior consumption of beef from a local abattoir. During investigation of the abattoir, bovine rectal swabs and samples of meat and surface swabs from beef carcasses were examined for E. coli 0157, isolates of which were tested for toxigenicity, plasmid content and phage type. E. coli 0157 was isolated from 84 (4%) of 2103 bovine rectal swabs; of these 84, 78 (93%) were VT', the most common phage types being 2 and 8, the types implicated in the cluster of human cases. Positive cattle were from diverse sources within England. E. coli 0157 was isolated from 7 (30%) of 23 carcasses of rectal swab-positive cattle and from 2 (8%) of 25 carcasses of rectal swab-negative cattle. The study has shown that cattle may be a reservoir of VT+ E. coli 0157, and that contamination of carcasses during slaughter and processing may be how beef and beef products become contaminated and thereby transmit the organism to man.

A dairy herd associated with Escherichia coli O157 infection in humans was studied for the 15 months following the outbreak to examine seasonal, age and management factors affecting faecal excretion of the organism and to determine the mode and frequency of milk contamination with the organism. Between May 1993 and July 1994, 28 visits were made to the farm to collect a total of 3593 rectal swabs from cows, heifers and calves and 329 milk samples. E. coli O157:H7 was isolated from 153 (4.3%) of 3593 bovine rectal swabs. The maximum prevalence at any one visit was 14% in lactating cows, 40% in non-lactating cows, 56% in calves and 68% in heifers. The prevalence in lactating cows, which was significantly lower than in the other groups, peaked during May-July 1993 and again briefly after the cattle were housed during November 1993 and then again during May 1994. Excretion rates of E. coli O157:H7 in lactating cows were highest during the first month after calving, falling during lactation and rising to another peak at 7 months postpartum. Between November 1993 and May 1994 there was no evidence of excretion in any group. Eighty-seven (74%) of the animals which excreted E. coli O157:H7 did so on only one occasion but 23 (32%) of 73 cows and heifers and 7 (16%) of 44 calves which excreted the organism did so on more than one occasion. E. coli O157:H7 was not isolated from milk taken from the bulk tank but it was isolated from individual milk samples (one milk jar and one fore-milk) from two animals previously shown to be faecal excretors of the organism. All isolates of E. coli O157:H7 obtained were of the same phage type, toxin genotype and plasmid profile.

Summary. Potassium tellurite was assessed for the selection of verocytotoxigenic (VT+) Escherichia coli 0 1 57. MICs were higher for VT+ E. coli 0 1 57 than for other strains of E. coli and for Aeromonas spp. MacConkey medium containing sorbitol, tellurite and cefixime (TC-SMAC) permitted the growth of VT+ E. coli 0157 and Shigella sonnei but partially or completely inhibited the growth of 67 O/ O of other strains of E. coli and all or most strains of other sorbitol-non-fermenting species tested. Of 39 1 rectal swabs from cattle screened on TC-SMAC medium, 26 yielded isolates of VT+ E. coli 0157 whereas sorbitol-MacConkey medium with cefixime and rhamnose yielded only nine isolates. Inclusion of potassium tellurite in sorbitol-MacConkey agar markedly increased the rate of isolation of VT+ E. coli 0157 from cattle rectal swabs and may do so for other types of specimen.

Summary. Enrichment culture (EC) in modified buffered peptone water followed by immunomagnetic separation (IMS) with magnetic beads coated with an antibody against Escherichia coli 0 157 (Dynabeads anti-E. coli 0 157 ; Dynal, Oslo) was compared with direct culture on cefixime rhamnose sorbitol MacConkey agar (CR-SMAC) and cefixime tellurite sorbitol MacConkey agar (CT-SMAC) for the isolation of E. coli 0157 from bovine faeces. When used to examine bovine faecal suspensions inoculated with 12 different strains of E. coli 0157, EC-IMS was c. 100-fold more sensitive for detection of the organism than direct culture on either medium. During monitoring of a dairy herd, E. coli 0 1 57 was isolated from 84 (8-2 YO) of 1024 rectal swabs taken from cattle over a 4-month period; 23 (27.4 YO of the 84 strains were isolated by both direct culture and IMS (1 5 of the 23 were isolated on both media, five on CT-SMAC only and three on CR-SMAC only), whereas 61 (72-6 YO) strains were isolated by IMS only. IMS is a sensitive and simple technique for the isolation of E. coli 0 157 from bovine faecal samples and should prove useful in elucidating further the epidemiology of this organism.